GSM7118777: WB, post-COVID-19, control MC1_96; Homo sapiens; RNA-Seq
Total RNA was extracted from 200 μL of preprocessed whole blood with the Maxwell® RSC simply RNA Blood Kit in combination with the Maxwell® RSC Instruments. RNA was isolated from 1 μg of total RNA using Illumina´s Ribo-zero plus rRNA depletion kit. The RNA was fragmented into approximately 200 base pair (bp) pieces by using divalent cations under elevated temperature. Reverse transcriptase and random primers were used to generate first strand cDNA from cleaved RNA fragments. DNA polymerase I and RNase H were used to obtain the second strand. Double-stranded cDNA fragments were end-repaired by Klenow DNA polymerase and T4 DNA.  cDNA was phosphorylated by T4 polynucleotide kinase and ligated to Illumina indexing adapters. Adapter-tagged libraries were amplified by using 15 cycles of PCR with DNA polymerase. Validation and quantification were performed using electrophoresis and qPCR. Pools of six indexed libraries were mixed at equimolar ratios to yield a total oligonucleotide mixture concentration of 10 nM.